DACE LAVOUT REFERENCES MAILINGS REVIEW VIEW DESIGN LAYOUT AaBbCr AaBbCet Aabl Aa

Question

DACE LAVOUT REFERENCES MAILINGS REVIEW VIEW DESIGN LAYOUT AaBbCr AaBbCet Aabl AaBbCet A 1Normal 11 No Spac," Heading 1 Heading 2 Title Subtitle So alx x, x' | A··a- | :, | :a . EE . Font Paragraph Styles TOTAAL: 50 Vraae 1/Question 1 U benodig n lise-buffer wat bestaan uit: U word voorsien van die volgende You need a lysis buffer consisting of the following: You are supplied with the following: 50 mM NaH PO 300 mM NaCl 75 mM Tris-Cl, pH 8 10% (v/v) glycerol 0.8 % sodium dodecyl sulfate 0.75 mg/ml lysonase NaH POs (Mr 119.98 g/mol) NaCI (Mr 58.443 g/mol) 2.5 M Tris-Cl, pH 8 oplossing / solution (1 Litre) 100 % glycerol 10 % Sodium dodecyl sulfate solution Lysonase (250 mg dissolved in 75 ml ddH:0) 12 How will you prepare 300 ml of the lysis buffer? Show all your steps and calculations. (2) Nac NaH PO 1000 mM NaH.POs is 119.98 in 1000 m/ water 1000 mM NaCI is 58.443 g in 1000 ml water

Explanation / Answer

Moles= grams/molar mass and Molarity= moles/volume

Given, Mr of NaH2PO4= 119.98 g/mol

119.98 g of NaH2PO4 can be dissolved in 1000ml of distilled water to make 1M or 1000mM NaH2PO4.

Let X ml of 1M NaH2PO4 is required to make 50mM of NaH2PO4 in 300ml lysis buffer.

Using the dilution formula,

M1V1=M2V2

1000xX= 50x300

X= 15ml

Given, Mr of NaCl=58.443 g/mol

58.443 g of NaCl can be dissolved in 1000ml of distilled water to make 1M or 1000mM NaCl.

Let X ml of 1M NaCl is required to make 300mM of NaClin 300ml lysis buffer.

Using the dilution formula,

M1V1=M2V2

1000xX= 300x300

X= 90ml

Using dilution formula, M1V1=M2V2

75x300= 2.5x1000xV2

V2= 75x300/2.5x1000

=9ml

9ml of 2.5M Tris-Cl (pH 8) is required to make 75mMTris-Cl (pH 8) in 300ml of lysis buffer.

Let X ml of 100% glycerol is required to prepare 10% glycerol in 300 ml of lysis buffer.

Therefore, 100xX = 10x300

X=30ml

30ml of 100% glycerol needs to be added to make up for 10% glycerol in 300ml of lysis buffer.

Let X ml of 10% SDS is required to prepare 0.8% SDS in 300 ml of lysis buffer.

Therefore, 10xX = 0.8x300

X=24ml

24ml of 10% SDS needs to be added to make up for 0.8% SDS in 300ml of lysis buffer.

Let X ml of 250mg/75ml Lysonase be required to make up 0.75mg/ml of it in the 300 ml of lysis buffer

Therefore, 0.75x300=250/75xX

X=67.5ml

A total of 235.5 ml of the above constituents and rest 64.5ml of distilled water will make up 300ml of the lysis buffer.

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